Team:HokkaidoU Japan/Notebook/September30

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{{Template:HokkaidoU_Japan}}
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*Digestion and Ligation of Arabinose Promoter,T3SSsignal and pSB1T3
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*Transformation
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<u>parts information</u>
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 +
Arabinose Promoter  1210bp(+primer:1259bp)
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 +
3-20B pSB2K3
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 +
 +
RBS+T3SSsignal  583bp(+primer:636bp)
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pSB1T3  2463bp(+primer:2504bp)
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== Digestion and Ligation of Arabinose Promoter,T3SSsignal and pSB1T3 ==
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{|style="text-align:center; float:left;" class="protocol"
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!Reagent
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!Amount
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|-
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|10x H buffer
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|2 uL
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|-
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|DW
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|1.6
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|-
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|pSB1T3
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|1
 +
|-
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|BSA
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|2
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|-
 +
|EcoR I
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|0.2
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|-
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|Pst I
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|0.2
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|-
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|style="border-top:1px solid #996;"|'''Total'''
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|style="border-top:1px solid #996;"|'''20 uL'''
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|}
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{|style="text-align:center; float:left;" class="protocol"
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!Reagent
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!Amount
 +
|-
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|10x H buffer
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|2 uL
 +
|-
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|DW
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|11.7
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|-
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|Promoter
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|3.5
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|-
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|BSA
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|2
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|-
 +
|EcoR I
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|0.5
 +
|-
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|Spe I
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|0.3
 +
|-
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|style="border-top:1px solid #996;"|'''Total'''
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|style="border-top:1px solid #996;"|'''20 uL'''
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|}
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{|style="text-align:center; float:left;" class="protocol"
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!Reagent
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!Amount
 +
|-
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|10x M buffer
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|5 uL
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|-
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|DW
 +
|32.8
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|-
 +
|T3SSsignal
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|2.5
 +
|-
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|BSA
 +
|5
 +
|-
 +
|Xba I
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|4.5
 +
|-
 +
|Pst I
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|0.2
 +
|-
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|style="border-top:1px solid #996;"|'''Total'''
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|style="border-top:1px solid #996;"|'''50 uL'''
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|}

Revision as of 07:20, 1 October 2010

Notebook
  • Digestion and Ligation of Arabinose Promoter,T3SSsignal and pSB1T3
  • Transformation


parts information

Arabinose Promoter 1210bp(+primer:1259bp)

3-20B pSB2K3


RBS+T3SSsignal 583bp(+primer:636bp)


pSB1T3 2463bp(+primer:2504bp)

Digestion and Ligation of Arabinose Promoter,T3SSsignal and pSB1T3

Reagent Amount
10x H buffer 2 uL
DW 1.6
pSB1T3 1
BSA 2
EcoR I 0.2
Pst I 0.2
Total 20 uL
Reagent Amount
10x H buffer 2 uL
DW 11.7
Promoter 3.5
BSA 2
EcoR I 0.5
Spe I 0.3
Total 20 uL
Reagent Amount
10x M buffer 5 uL
DW 32.8
T3SSsignal 2.5
BSA 5
Xba I 4.5
Pst I 0.2
Total 50 uL
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/September30"